11 alpha-hydroxy-4-androstene-3, 17-dione



Patented Oct. 20, 1953 Herrera d nimray; Hickory earners", aminutty H. P erso n, Kalamazoo, Mich., assignors to oh n' (Joirifiany, Kala-nfa'zdo, Mich; corporation of Michigan No Drawing. Aii'fiiicatiof 19', 1952, Seriaf No. 277,523-

This inventionrelates to a new compound, 11anydroxyaridrosteaeuiefie.

An object of this invention is to provide 11a.- hydroxyandrostne'dione having pharmacological activity. A further object is to prepare an intermediate useful in the synthesis of known 11- oxygenated steroids. The lla-hYdIOXYELlldIO- stenedione may be converted to the known adrenosterone (4 androstene 3,11,17 trione) by oxidation.

11a-hydroxyandrostenedione may be prepared from androstenedione as described in our applications, of which this is a continuation-in-part, Serial No. 180,496, filed August 19, 1950, and Serial No. 272,944, filed February 23, 1952.

The 1la-hydroxy-4-androstene-3,17-dione has unusual and desirable physical and pharmacological properties. It demonstrates anaesthetic and inhibitory properties in estrogen and leuteoid activities. The thermal stability of the compound renders it particularly useful in reactions and purification techniques at elevated temperature. The susceptibility of the compound to acylation permits protection of the hydroxyl group in oxidation procedures.

The following examples are illustrative of the processes and products of the present invention and are not to be construed as limiting.

Example 1.11a-hydr0my-4-androstene- 3,1 7 -dine time the fermentation liquor and mycelia were extracted suocesssively with three one-liter portions, one two-liter portion, and one one-liter portion of methylene chloride. The methylene chloride extracts were combined and washed with two 100-milliliter portions of two percent aqueous sodium bicarbonate solution and three 500- milliliter portions of distilled water. The methylene chloride extract was evaporated to dryness in vacuo and the solids were taken up in fifty milliliters of methylene chloride. The solution was transferred to a 100-milliliter beaker and evap- 1 Glainr. (Cl. 260-397-4) onrsmatograpmc separation over an Ali'Qs packed column yielded three fractions, one of which was 11d=1 I'-yfi I-'G1'iy androstenedione'.

Example 2.11a-hydroxy-4-androstene- 3,17-dione of Cooper, Fernstrom and Miller, Ind. Eng. Chem,

36, 504 (1944). To this medium containing a 24 hour growth of Rhizopus arrhizus was added one gram of 4-androstene-3,17-dione in 25 milliliters of acetone to provide a suspension of the steroid in the culture. After an additional 24 hour period of incubation under the same conditions of temperature and aeration, the beer and mycelium were extracted. The mycelium was filtered, washed twice, each time with a volume of acetone approximately equal to the volume of the mycelium and extracted twice, each time with a volume of methylene chloride approximately equal to the volume of the mycelium. The acetone and methylene chloride extracts including solvent were added to the beer filtrate. The mixed extracts and beer filtrate were extracted successively with two one-half by volume portions of methylene chloride and then with two one-fourth by volume portions of methylene chloride. The combined methylene chloride extracts were washed with two one-tenth by volume portions of a two percent aqueous solution of sodium bicarbonate and then with two one-tenth by volume portions of water. After drying the methylene chloride with about three to five grams of anhydrous sulfate per liter of solvent and filtering, the solvent was removed by distillation. The extract of 2.09 grams was dissolved in milliliters of benzene and chromatographed over fifty grams of hydrochloric acid washed, degrees centigrade heated alumina with 100 milliliter portions of developing solvent, as indicated in the table.

The crystalline fractions 18 to 21 inclusive were washed with several milliliters of ether. The residue was recrystallized from ten milliliters of ethyl acetate, adding Skellysolve B petroleum,

naphtha to effect crystallization. After four crystallizations, 240 milligrams of 11a-hydroxy-4- androstene-3,l7-dione were obtained, melting at 226 to 227 degrees centigrade. The structure was verified by infrared, ultraviolet spectrum, and oxidation with chromic acid-glacial acetic acid to adrenosterone. Reichstein, Helv. Chim. Acta, 20, 987 (1937) has prepared a different isomer, the 115-hydroxy-4-androstene-3,17-dione melting at 189 to 190 degrees centigrade. The Ila-hYdIOXY- 4-androstene-3,17-dione has an optical rotation [111 of plus 162 degrees (1.415 in chloroform) and a km of 50.54.

Analysis: Calculated for CHI-I26 03: C, 75.46; H, 8.66. Found: C, 75.68; H, 8.53.

Fractions 16 and 17 combined and recrystallized three times from acetone by the addition of Skellysolve B yielded a. compound melting at 195 to 196 degrees centigrade, analyzing as C19H2603 and showing no absorption in the ultraviolet spectrum. Infrared spectrum showed the presence of three ketonic groups.

Analysis: Calculated for Ciel-12603: C, 75.46; H, 8.66. Found: C, 75.73; H, 8.68.

TABLE Eluate Fraction Solvent Solids,

Milligrams hpnmne 322 benzene plus 5 percent ether 108 benzene plus 10 percent ethen. 10 venzene plus percent ether- 12 ether 15 ether plus 5 percent 011013 19 ether plus 10 percent GHC13 23 ether plus 50 percent GHOl; chloroform 716 CHO]; plus 5 percent aceton 177 CHC-h plus 10 percent acetone 71 011013 plus 50 percent acetone 26 acetone 41 acetone plus 5 percent methanoL acetone plus 10 percent methanol 42 acetone plus 50 percent methanol 40 methanol 13 We claim:

l1a-hydroxy-4-androstene-3,17-dione.

HERBERT C. MURRAY. DUREY H. PETERSON.

No references cited. 

